Establishment and characterization of a novel in vitro angiogenesis model using a microvascular endothelial cell line, F-2C, cultured in chemically defined medium.
The behavior of vascular endothelial cells (EC) is an important factor in the processes involved in angiogenesis, but the regulatory mechanisms of angiogenesis, especially underlying the tubulogenesis by EC are not yet clear. Although a number of in vitro experimental models of tubulogenesis have been developed by use of cultured EC, most of those models are too complex to be easily handled and further, the culture media are usually supplemented with serum, creating problems in interpretation of experimental results. To generate a simple in vitro angiogenesis study model under serum-free culture conditions, we adapted a murine microvascular endothelial cell line, F-2, to a chemically defined medium, Cos Medium 001, and successfully established a subline of F-2, designated F-2C, which revealed a unique growth pattern. In Cos Medium 001, F-2C proliferates in a cobblestone pattern at an early growth stage, but, at a late growth stage, spontaneously differentiates to form three-dimensional honeycomblike tubular structures without the supplementation of any specific factors. The cell aggregation activity of F-2C in the presence of Ca2+ was much greater than that of F-2. The amount of subendothelial matrix deposited by F-2C was significantly higher than that by F-2, and increased prominently after the F-2C cells reached the differentiating stage of tubulogenesis. These findings indicate that F-2C is a new EC line in which tubulogenesis is spontaneously induced by the marked deposition of basement membrane analog to the subendothelial matrix and by the enhancement of presumable cadherin activity. We suggest that this cell line, F-2C, represents a simple and useful in vitro angiogenesis model.
Chen
C S
CS
Department of Dermatology, Faculty of Medicine, Kyoto University, Japan.
Toda
K I
KI
Maruguchi
Y
Y
Matsuyoshi
N
N
Horiguchi
Y
Y
Imamura
S
S
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002448:Cell Adhesion; D002449:Cell Aggregation; D002454:Cell Differentiation; D002455:Cell Division; D002460:Cell Line; D016895:Culture Media, Serum-Free; D004730:Endothelium, Vascular / Q000166:cytology*; D005109:Extracellular Matrix / Q000502:physiology; D051379:Mice; D008833:Microcirculation / Q000033:anatomy & histology; D008954:Models, Biological; D018919:Neovascularization, Physiologic
D016895:Culture Media, Serum-Free; D002118:Calcium